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Drought adaptations and responses in five genotypes of Fraxinus pennsylvanica Marsh.: photosynthesis, water relations and leaf morphology.

Genotypic variation in photosynthesis and plant water relations during drought, and in leaf and seedling morphology were examined in greenhouse-grown Fraxinus pennsylvanica Marsh. (green ash) from five populations located along an east-west transect from New York State to South Dakota. During a 17-day drought, South Dakota seedlings, from the most xeric habitat, maintained the highest net photosynthesis and leaf conductance, and New York seedlings, from the most mesic habitat, exhibited the lowest net photosynthesis and leaf conductance. All populations except New York adjusted osmotically during the 17-day drought, by the end of which New York seedlings had the highest osmotic potentials at full and zero turgor. Tissue elasticity increased in New York seedlings, but decreased in Nebraska seedlings during the drought. Leaves of South Dakota seedlings were the most xerophytic. They were smaller in area and greater in thickness and specific mass than leaves of other sources. Leaves of New York seedlings were thinner than those of the other genotypes and among the largest. Seedlings from South Dakota were smaller than those of the other populations.

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Degree of C(4) Photosynthesis in C(4) and C(3)-C(4)Flaveria Species and Their Hybrids : I. CO(2) Assimilation and Metabolism and Activities of Phosphoenolpyruvate Carboxylase and NADP-Malic Enzyme.

The degree of C(4) photosynthesis was assessed in four hybrids among C(4), C(4)-like, and C(3)-C(4) species in the genus Flaveria using (14)C labeling, CO(2) exchange, (13)C discrimination, and C(4) enzyme activities. The hybrids incorporated from 57 to 88% of the (14)C assimilated in a 10-s exposure into C(4) acids compared with 26% for the C(3)-C(4) species Flaveria linearis, 91% for the C(4) species Flaveria trinervia, and 87% for the C(4)-like Flaveria brownii. Those plants with high percentages of (14)C initially fixed into C(4) acids also metabolized the C(4) acids quickly, and the percentage of (14)C in 3-phosphoglyceric acid plus sugar phosphates increased for at least a 30-s exposure to (12)CO(2). This indicated a high degree of coordination between the carbon accumulation and reduction phases of the C(4) and C(3) cycles. Synthesis and metabolism of C(4) acids by the species and their hybrids were highly and linearly correlated with discrimination against (13)C. The relationship of (13)C discrimination or (14)C metabolism to O(2) inhibition of photosynthesis was curvilinear, changing more rapidly at C(4)-like values of (14)C metabolism and (13)C discrimination. Incorporation of initial (14)C into C(4) acids showed a biphasic increase with increased activities of phosphoenolpyruvate carboxylase and NADP-malic enzyme (steep at low activities), but turnover of C(4) acids was linearly related to NADP-malic enzyme activity. Several other traits were closely related to the in vitro activity of NADP-malic enzyme but not phosphoenolpyruvate carboxylase. The data indicate that the hybrids have variable degrees of C(4) photosynthesis but that the carbon accumulation and reduction portions of the C(4) and C(3) cycles are well coordinated.

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Energy storage of linear and cyclic electron flows in photosynthesis.

The energy storage of photosynthesis in the green alga Chlorella vulgaris was determined by pulsed, time-resolved photoacoustics. The energy storage of the linear electron transfer process in photosynthesis, of cyclic photosystem (PS) I, and possibly of PSII was determined by selection of excitation wavelength and of flash interval. At 695 nm excitation, a rather large cyclic PSI energy storage of 0.68 +/- 0.04 eV/quantum of energy at 8 ms after a 1-mus flash was obtained. This energy remained the same at flash intervals of 0.35 to 60 s and was independent of the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. We tentatively assign this energy to the ferredoxin-NADP-reductase-ferredoxin and oxidized cytochrome b(6)/f complexes. An efficient distribution of energy between cyclic and linear systems is obtained with the simple assumption that the turnover time of the cyclic system is slower than that of the linear system. The energy storage of linear electron flow was determined by 655 nm excitation of Chlorella with a short flash interval of 0.35 s per flash. It was calculated to be 0.50 +/- 0.03 eV/hv, close to that expected for oxygen and NADPH formation. The energy storage of PSII is determined by excitation of Chlorella at 655 nm with a long flash interval of 60 s per flash. It was calculated to be 1.07 +/- 0.05 eV/hv, consistent with the energy storage being in S-states and the secondary electron acceptor of PSII with a calculated redox energy of 1.03 eV/hv. In the presence of 1 mum 3-(3,4-dichlorophenyl)-1,1-dimethylurea, the calculated energy storage in PSII is still significant, 0.53 +/- 0.04 eV/hv. This probably indicates a significant cyclic electron flow around PSII. These cyclic flows may contribute considerably to energy storage in photosynthesis.

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Regulatory Phosphorylation of C4 Phosphoenolpyruvate Carboxylase (A Cardinal Event Influencing the Photosynthesis Rate in Sorghum and Maize).

C4 leaf phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) is subject to a day/night regulatory phosphorylation cycle. By using the cytoplasmic protein synthesis inhibitor cycloheximide (CHX), we previously reported that the reversible in vivo light activation of the C4 PEPC protein-serine kinase requires protein synthesis. In the present leaf gas-exchange study, we have examined how and to what extent the CHX-induced inhibition of PEPC protein kinase activity/PEPC phosphorylation in the light influences C4 photosynthesis. Detached Sorghum vulgare and maize (Zea mays) leaves fed 10 [mu]M CHX showed a gradual but marked decrease in photosynthetic CO2 assimilation capacity. A series of control experiments designed to assess deleterious secondary effects of the inhibitor established that this reduction in C4 leaf CO2 assimilation was not due to (a) an increased stomatal resistance to CO2 diffusion, (b) a decrease in the activation state of other photoactivated C4 cycle enzymes, and (c) a perturbation of the Benson-Calvin C3 cycle, as evidenced by the absence of an inhibitory effect of CHX on leaf photosynthesis by a C3 grass (Triticum aestivum). It is notable that the CHX-induced decrease in CO2 assimilation by illuminated Sorghum leaves was highly correlated with a decrease in the apparent phosphorylation status of PEPC and a concomitant change in carbon isotope discrimination consistent with a shift from a C4 to a C3 mode of leaf CO2 fixation. These collective findings indicate that the light-dependent activation of the PEPC protein-serine kinase and the resulting phosphorylation of serine-8 or serine-15 in Sorghum or maize PEPC, respectively, are fundamental regulatory events that influence leaf C4 photosynthesis in vivo.

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Measurement of the Leakage of CO2 from Bundle-Sheath Cells of Leaves during C4 Photosynthesis.

During C4 photosynthesis, CO2 is released in bundle-sheath cells by decarboxylation of C4 acids and then refixed via ribulose-1,5-bisphosphate carboxylase. In this study we examined the efficiency of this process by determining the proportion of the released CO2 that diffuses back to mesophyll cells instead of being refixed. This leak of CO2 was assessed by determining the amount of 14CO2 released from leaves during a chase in high [12CO2] following a 70-s pulse in 14CO2. A computer-based analysis of the time-course curve for 14CO2 release indicated a first-order process and provided an estimate of the initial velocity of 14CO2 release from leaves. From this value and the net rate of photosynthesis determined from the 14CO2 fixed in the pulse, the CO2 leak rate from bundle-sheath cells (expressed as a percentage of the rate of CO2 production from C4 acids) could be deduced. For nine species of Gramineae representing the different subgroups of C4 plants and two NAD-malic enzyme-type dicotyledonous species, the CO2 leak ranged between 8 and 14%. However, very high CO2 leak rates (averaging about 27%) were recorded for two NADP-malic enzyme-type dicotyledonous species of Flaveria. The results are discussed in terms of the efficiency of C4 photosynthesis and observed quantum yields.

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Modification of Photosystem I Light Harvesting of Bundle-Sheath Chloroplasts Occurred during the Evolution of NADP-Malic Enzyme C4 Photosynthesis.

Low-temperature emission spectra and excitation spectra for chlorophyll fluorescence were recorded from leaves of species of the genus Flaveria (Asteraceae) with C3, C3-C4-intermediate, C4-like, and C4 photosynthesis. Among the latter two groups, high chlorophyll b absorption was observed in excitation spectra for photosystem I (PSI) fluorescence. By comparing leaf data with those from isolated chloroplast fractions, the high chlorophyll b absorption was attributed to the specific properties of the bundle-sheath chloroplasts in leaves from C4 plants. The deconvolution of the PSI excitation spectra and the use of a model revealed that the contribution of photosystem II absorption to the functional antenna of PSI was markedly increased in leaves from three of the five C4-like and C4 species investigated in detail. The two other species exhibited normal, C3-like light-harvesting properties of PSI. The former species are known for efficient carbon assimilation, the latter for decreased efficiencies of carbon assimilation. It is concluded that photosystem II becomes a substantial part of the functional PSI antenna late in the evolution of C4 photosynthesis, and that the composite antenna optimizes the light-harvesting of PSI in bundle-sheath chloroplasts to meet the energy requirements of C4 photosynthesis.

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Acclimation of photosynthesis to elevated CO2 under low-nitrogen nutrition is affected by the capacity for assimilate utilization. Perennial ryegrass under free-Air CO2 enrichment

Acclimation of photosynthesis to elevated CO2 has previously been shown to be more pronounced when N supply is poor. Is this a direct effect of N or an indirect effect of N by limiting the development of sinks for photoassimilate? This question was tested by growing a perennial ryegrass (Lolium perenne) in the field under elevated (60 Pa) and current (36 Pa) partial pressures of CO2 (pCO2) at low and high levels of N fertilization. Cutting of this herbage crop at 4- to 8-week intervals removed about 80% of the canopy, therefore decreasing the ratio of photosynthetic area to sinks for photoassimilate. Leaf photosynthesis, in vivo carboxylation capacity, carbohydrate, N, ribulose-1,5-bisphosphate carboxylase/oxygenase, sedoheptulose-1,7-bisphosphatase, and chloroplastic fructose-1, 6-bisphosphatase levels were determined for mature lamina during two consecutive summers. Just before the cut, when the canopy was relatively large, growth at elevated pCO2 and low N resulted in significant decreases in carboxylation capacity and the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase protein. In high N there were no significant decreases in carboxylation capacity or proteins, but chloroplastic fructose-1,6-bisphosphatase protein levels increased significantly. Elevated pCO2 resulted in a marked and significant increase in leaf carbohydrate content at low N, but had no effect at high N. This acclimation at low N was absent after the harvest, when the canopy size was small. These results suggest that acclimation under low N is caused by limitation of sink development rather than being a direct effect of N supply on photosynthesis.

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Interactions between senescence and leaf orientation determine in situ patterns of photosynthesis and photoinhibition in field-grown rice

Photosynthesis and photoinhibition in field-grown rice (Oryza sativa L.) were examined in relation to leaf age and orientation. Two varieties (IR72 and IR65598-112-2 [BSI206]) were grown in the field in the Philippines during the dry season under highly irrigated, well-fertilized conditions. Flag leaves were examined 60 and 100 d after transplanting. Because of the upright nature of 60-d-old rice leaves, patterns of photosynthesis were determined by solar movements: light falling on the exposed surface in the morning, a low incident angle of irradiance at midday, and light striking the opposite side of the leaf blade in the afternoon. There was an early morning burst of CO2 assimilation and high levels of saturation of photosystem II electron transfer as incident irradiance reached a maximum level. However, by midday the photochemical efficiency increased again almost to maximum. Leaves that were 100 d old possessed a more horizontal orientation and were found to suffer greater levels of photoinhibition than younger leaves, and this was accompanied by increases in the de-epoxidation state of the xanthophyll cycle. Older leaves had significantly lower chlorophyll content but only slightly diminished photosynthesis capacity.

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Extracellular carbonic anhydrase facilitates carbon dioxide availability for photosynthesis in the marine dinoflagellate prorocentrum micans

This study investigated inorganic carbon accumulation in relation to photosynthesis in the marine dinoflagellate Prorocentrum micans. Measurement of the internal inorganic carbon pool showed a 10-fold accumulation in relation to external dissolved inorganic carbon (DIC). Dextran-bound sulfonamide (DBS), which inhibited extracellular carbonic anhydrase, caused more than 95% inhibition of DIC accumulation and photosynthesis. We used real-time imaging of living cells with confocal laser scanning microscopy and a fluorescent pH indicator dye to measure transient pH changes in relation to inorganic carbon availability. When steady-state photosynthesizing cells were DIC limited, the chloroplast pH decreased from 8.3 to 6.9 and cytosolic pH decreased from 7.7 to 7.1. Re-addition of HCO3- led to a rapid re-establishment of the steady-state pH values abolished by DBS. The addition of DBS to photosynthesizing cells under steady-state conditions resulted in a transient increase in intracellular pH, with photosynthesis maintained for 6 s, the amount of time needed for depletion of the intracellular inorganic carbon pool. These results demonstrate the key role of extracellular carbonic anhydrase in facilitating the availability of CO2 at the exofacial surface of the plasma membrane necessary to maintain the photosynthetic rate. The need for a CO2-concentrating mechanism at ambient CO2 concentrations may reflect the difference in the specificity factor of ribulose-1,5 bisphosphate carboxylase/oxygenase in dinoflagellates compared with other algal phyla.

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Seasonal Course of Photosynthesis, Respiration, and Distribution of C in Young Pinus resinosa Trees as Related to Wood Formation.

Rates of net photosynthesis and dark respiration, and distribution of (14)C were determined for new (current season's) and old (previous season's) needles at 10 times during the seasonal development of young Pinus resinosa Ait. trees. The seasonal changes in these factors associated with the development of the new shoot were related to known seasonal patterns of wood formation.Net photosynthesis per gram of needle dry weight (photosynthetic efficiency) was maximum in the old needles at the time of first new needle elongation; at the same time translocation of (14)C from old to new needles was greatest. Photosynthetic efficiency of new needles was maximum at the end of the period of rapid new needle elongation, when the new needles also began exporting much greater quantities of (14)C to other plant parts. In particular, the amount translocated from the new needles to the stem was greatly increased. At this time thick-walled xylem cells were first observed in the stem.These results, together with those of previous studies, indicate that the production of thick-walled xylem tracheids normally associated with latewood is physiologically correlated with maturation of the current season's needles. Because there is a lesser demand for photosynthate in the new shoot and a high rate of photosynthesis in the whole plant at the time of new needle maturity, a sharply increased amount of photosynthate becomes available for wall synthesis by cambial derivatives.

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Variation in photorespiration. The effect of genetic differences in photorespiration on net photosynthesis in tobacco.

The hypothesis that net photosynthesis is diminished in many plant species because of a high rate of CO(2) evolution in the light has been tested further. High rates of CO(2) output in CO(2)-free air in comparison with dark respiration were found in Chlamydomonas reinhardi, wheat leaves, tomato leaves, and to a lesser extent in Chlorella pyrenoidosa by means of the (14)C-photorespiration assay. In tobacco leaves high photorespiration was characteristic of a standard variety, Havana Seed, and a possibly still higher rate was found in a yellow heterozygous mutant, JWB Mutant. However, the dark homozygous sibling of the latter, JWB Wild, had a low photorespiration for the tobacco species. The relative rates of photorespiration were in the same sequence when measured by the (14)CO(2) released in normal air from leaf disks supplied with glycolate-1-(14)C in the light.As would be predicted by the hypothesis, the maximal net rate of photosynthesis at 300 ppm of CO(2) in the air in JWB Wild leaves was greater (24%) than in Havana Seed, while JWB Mutant had less CO(2) uptake than the standard variety (21%). At 550 ppm of CO(2) the differences in net photosynthesis were not as great between the 2 siblings as at 200 ppm. The relative leaf expansion rates of seedlings of the 3 tobacco varieties in a greenhouse had the same relationship as their rates of CO(2) assimilation.Thus within the tobacco species, as in a comparison between tobacco and maize, low photorespiratory CO(2) evolution was correlated with higher photosynthetic efficiency. Therefore it seems that increased CO(2) uptake should be achieved by genetic interference with the process of photorespiration.

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Decrease in net photosynthesis caused by respiration.

CO(2) exchange between air and leaf is conceived as a current along a resistor with a respiratory current, unaffected by CO(2) concentration, entering the resistor. The conclusion follows that a plant placed in an atmosphere free of CO(2) will increase the CO(2) concentration to the compensation concentration along a curve determined by the resistor and the volume of the atmosphere. This was verified. Also a photosynthesis rate calculated from the parameters of the observed curve agreed with an independent observation of photosynthesis in CO(2)-free air. The decrease in net photosynthesis caused by respiration is, according to the model, the CO(2) compensation concentration divided by the concentration in the atmosphere.

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Studies on electron-transport reactions of photosynthesis in plastome mutants of oenothera.

Fluorescence characteristics and light-induced absorbance changes of 5 plastome mutants of Oenothera, all having a defect in photosynthesis, were investigated to localize the site of the block in their photosynthetic mechanism and to relate mutational changes in the plastome to specific biochemical events in photosynthesis. In 4 of the mutants examined photosystem 2 was largely, or completely, nonfunctional. Excitation of system 2 did not cause reduction of oxidized cytochrome f in these mutants. The system-2 dependent absorbance change at 518 mmu seen in normal leaves was absent in the mutants. Moreover, the mutants had a high initial fluorescence in the presence and in the absence of 3- (3,4-dichlorophenyl)-1,1-dimethylurea, which did not change during illumination, indicating that the reaction centers of system 2 were affected by the mutations. Photosystem 1 functioned normally.A fifth mutant had an impaired photosystem 1. Even high intensity far-red light did not lead to an accumulation of oxidized cytochrome f as was seen in normal plants. Photosystem 2 was functioning, as evidenced by the fast reduction of the primary system-2 oxidant, and by the characteristics of the 518-mmu absorbance change.Because 1 of the 2 photosystems is functional in all mutants, and because they all have the enzymes of the photosynthetic carbon cycle, it appears that the effect of the mutation is specific. The results suggest that the plastome controls reactions within the electron-transport chain of photosynthesis.

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Photosynthesis, Photorespiration and Respiration of Chloroplasts From Acetabularia mediterrania.

A chloroplast fraction isolated from Acetabularia mediterrania carries on photosynthesis at rates essentially equal to those of whole cells. Electron and phase contrast microscopy reveals that the chloroplasts are intact and well preserved. Preparations contain no identifiable peroxisomes, but some cytoplasmic and mitochondrial contamination is present. Photosynthesis and CO(2) production in light by chloroplast preparations are in many respects similar to that of bean leaves, although the measured rates are somewhat lower. Respiration and photosynthesis of chloroplast preparations and whole cells of Acetabularia is essentially similar except that cells have a strong dark-type respiration which continues in light and is CO(2) dependent, the substrate being mainly recent photosynthate. The data suggest that chloroplasts are the site of photorespiration.

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The effects of simazine and temperature on photosynthesis in rye.

Temperature during growth had a strong influence on light-saturated rates of photosynthesis and respiration in rye (Secale cereale). Carbon dioxide uptake was depressed at the low temperatures and reached maximal values higher than 30 milligrams of carbon dioxide per square decimeter per hour at the intermediate temperatures. Respiration rates increased substantially while growth temperature decreased. Simazine treatment (0.00, 0.03, 0.08, 0.12 milligrams per liter) resulted in no significant stimulatory or inhibitory effects at low temperatures, but progressively inhibited growth and photosynthesis at the higher temperatures. Respiration rates were not significantly influenced. Thus, the effect of simazine is strongly temperature-dependent and is more pronounced on photosynthesis than respiration.

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Carbon dioxide compensation points of leaves and stems and their relation to net photosynthesis.

The interactions between CO(2) and H(2)O vapor exchange of the leaf and respirant organs like stems were studied in tobacco plants. The results were analyzed according to a suggested model. Good agreement between open and closed system measurements supported the validity of the model.The measured over-all resistance to CO(2) of a leaf and a stem enclosed in a measuring cuvette was the same as the measured resistance of the leaf when measured alone provided the resistance of the stem to CO(2) is relatively high. The combined CO(2) compensation concentration of a leaf and stem having high resistance to CO(2) was higher than the CO(2) compensation point of the leaf alone, by the magnitude of rate of CO(2) evolution from the stem multiplied by the overall resistance of the leaf.CO(2) evolution into CO(2)-free air was found to be higher in light than in dark in leaves, while the reverse was true for stems. It was concluded that normally the CO(2) compensation point of a leaf is unaffected by stomata and boundary layer resistance while the combined CO(2) compensation point of a leaf and a stem differs in its nature since it represents a steady state of photosynthesis in which stem contribution, I(a), is equal to net photosynthesis, I(s). Interpretation of the experimental data shows tht respiration in the light is unaffected by external CO(2) concentration (at the range of 0-300 mul liter) and by intensity of photosynthesis.

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Is nicotinamide adenine dinucleotide phosphate an obligatory intermediate in photosynthesis?

The site of action of the inhibitors disalicylidenepropanediamine and pyrophosphate was more closely defined as acting on ferredoxin. Three inhibitors which act on the electron transport path between ferredoxin and NADP: disalicylidenepropanediamine, pyrophosphate, and phosphoadenosinediphosphate ribose, had no effect on photosynthesis in cell free preparations of Dunaliela parva at concentrations which completely inhibited the enzymic activity on which each inhibitor acts. The addition of disalicylidenepropanediamine to dark-grown Euglena gracilis cells prevented the light-induced formation of NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, but not of photosynthesis, chlorophyll synthesis, or NAD-dependent glyceraldehyde-3-phosphate dehydrogenase.The above results are interpreted as indicating that, at least under some conditions, a reduced product of photosystem I preceding ferredoxin in the electron transport path can serve as the reductant of CO(2) in photosynthesis.

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Changes in respiration, photosynthesis, adenosine 5'-triphosphate, and total adenylate content of ozonated pinto bean foliage as they relate to symptom expression.

The effect of 0.25 to 0.30 microliter per liter ozone on photosynthesis and respiration and on the ATP and total adenylate content of the primary leaves of pinto beans (Phaseolus vulgaris L.) was examined. Changes in these parameters over a 72-hour time period were correlated with the development of symptoms of ozone toxicity. Toxicity symptoms normally appeared within 24 hours. The content of ATP and total adenylates increased immediately following a 3-hour exposure to ozone. Photosynthesis was depressed initially, but returned to normal within 24 hours. Respiration was not always altered initially, but it was significantly stimulated within 24 hours. We interpret the results to mean that the changes in adenylate content and photosynthesis are early events in the initiation of ozone damage and that the change in respiration is a consequence rather than a cause of cellular injury.

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